English  |  正體中文  |  简体中文  |  Items with full text/Total items : 27533/39387
Visitors : 2537912      Online Users : 27
RC Version 4.0 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
Scope Adv. Search
LoginUploadHelpAboutAdminister

Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/49021

Title: 以小鼠巨噬細胞 RAW 264.7 細胞模式探討幾丁聚醣分子量對抗發炎活性之影響
Effect of molecular weight of chitosan on anti-inflammatory activities using macrophage RAW 264.7 model
Authors: Lin, Yi-Yung
林奕雍
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 幾丁聚醣;小鼠巨噬細胞;發炎;細胞激素;MAPK
chitosan;RAW264.7 macrophages;inflammation;cytokines;MAPK
Date: 2016
Issue Date: 2018-08-22T06:40:54Z
Abstract: 本研究主要探討幾丁聚醣分子量大小對其抗發炎活性之影響。分子量 300 kDa 幾丁聚醣經纖維素酶水解及甲醇區分,得到分子量分別為 156、72、29.2、7.1、3.3、2.2 kDa 等六種不同分子量幾丁聚醣降解產物及水解 18 小時所得幾丁寡醣,取這八種不同分子量幾丁聚醣及幾丁寡醣為測試樣品,評估抗發炎的潛力。結果顯示分子量大於 29.2 kDa (300、156 及 72 kDa) 的幾丁聚醣均能抑制 LPS (Lipopolysaccharide) 誘導細胞產生 NO (Nitrite oxide),其中 156 及 72 kDa 於 250 μg/mL 可達 84.39% 與 75.67% 抑制率;而 ≦ 29.2 kDa 的幾丁聚醣均有促發炎的活性,250 μg/mL 的 7.1 kDa 與 18 hr COS (18 hr chitooligosaccharides) 使細胞 NO 分泌量增加 17.30% 及 12.66%。156 與 72 kDa 皆能顯著降低促發炎激素 TNF-α (Tumor necrosis factor alpha) 及 IL-6 (Interleukin-6) 的含量,且具濃度依存性;7.1 kDa 及 18 hr COS 於 250 μg/mL 可顯著提高 TNF-α 及 IL-6 含量。而在抗發炎激素 IL-10 (Interleukin-10) 皆隨著劑量上升而提高,但透過 IL-6/IL-10 比值的結果發現 156 與 72 kDa 最具抗發炎活性。以 Western blot 分析幾丁聚醣對 RAW264.7 細胞分泌 NO 之訊息傳遞路徑,156 與 72 kDa 可藉由抑制 MAPK (Mitogen-activated protein kinases) 中 ERK、JNK 及 p38 訊息蛋白的活化,進而降低 NF-κB 活性及 iNOS (Inducible NOS) 蛋白表現量,減少巨噬細胞分泌 NO;7.1 kDa 與 18 hr COS 則是以活化 MAPK 中 JNK 訊息蛋白,活化 NF-κB 及 iNOS 蛋白表現量,進而增加 NO 分泌量。最後,分別以 anti-CD14、anti-TLR4 及 anti-CR3 抗體處理細胞,發現 156 kDa 是由 CR3 受器抑制 RAW264.7 細胞產生 NO,72 kDa 藉由 TLR4 及 CR3 受器抑制細胞產生 NO,7.1 kDa 及 18 hr COS 則是透過 CD14、TLR4 及 CR3 等受器調節 NO 生成。
The effect of chitosan with various molecular weight on anti-inflammatory activity was evaluated. Chitosan samples with molecular weights (MWs) of 156, 72, 29.2, 7.1, 3.3, 2.2 kDa and chitooligosaccharides (COS) were prepared by cellulase degradation of a 300 kDa chitosan, followed by methanol fractionation. The effects of these chitosan products on NO (Nitric oxide), cytokines production and involved MAPK (Mitogen-activated protein kinases) signal transduction pathway in LPS (Lipopolysaccharide)-treated murine macrophage RAW264.7 were investigated. Chitosans with MWs > 29.2 kDa (300, 156, 72 kDa) had the better inhibition on NO production, among which 156 and 72 kDa chitosans were the highest with 84.39% and 75.67% inhibition at 250 μg/mL; whereas chitosans with MWs ≦ 29.2 kDa had pro-inflammation effect, samples of 7.1 kDa chitosan and 18 hr COS significantly increase NO production by 17.30% and 12.66%, respectively. The 156 and 72 kDa chitosans significantly inhibited pro-inflammatory cytokines production, such as TNF-α (Tumor necrosis factor alpha) and IL-6 (Interleukin-6), in LPS-treated RAW264.7 macrophages in a dose-dependent manner, whereas 7.1 kDa chitosan and 18 hr COS signification induced TNF-α and IL-6 production. Although IL-10 production was increased with the increasing of the dosage for all test samples, the 156 and 72 kDa chitosans had the strongest anti-inflammatory potential by the highest decreasing the ratio of IL-6/IL-10. The 156 and 72 kDa chitosans via binding with the receptor CR3 could inhibit the activation of the signal proteins of ERK, JNK and p38, then further inhibit the NF-κB activation and iNOS expression, as evidenced by western blot analysis. In the other hand, the 7.1 kDa chitosan and 18 hr COS via binding with the receptors of CD14, TLR4 and CR3 to activate JNK phosphorylation, then, to activate NF-κB and enhance iNOS expression, and accordingly, to increase NO production.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010332046.id
http://ntour.ntou.edu.tw:8080/ir/handle/987654321/49021
Appears in Collections:[食品科學系] 博碩士論文

Files in This Item:

File Description SizeFormat
index.html0KbHTML11View/Open


All items in NTOUR are protected by copyright, with all rights reserved.

 


著作權政策宣告: 本網站之內容為國立臺灣海洋大學所收錄之機構典藏,無償提供學術研究與公眾教育等公益性使用,請合理使用本網站之內容,以尊重著作權人之權益。
網站維護: 海大圖資處 圖書系統組
DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - Feedback